Mebendazole Treatment Disrupts the Transcriptional Activity of Hypoxia-Inducible Factors 1 and 2 in Breast Cancer Cells

Natalie S Joe; Yuanfeng Wang; Harsh H Oza; Inês Godet; Nubaira Milki; Gregory J Riggins; Daniele M Gilkes

doi:10.3390/cancers15041330

Mebendazole Treatment Disrupts the Transcriptional Activity of Hypoxia-Inducible Factors 1 and 2 in Breast Cancer Cells

Cancers (Basel). 2023 Feb 20;15(4):1330. doi: 10.3390/cancers15041330.

Authors

Natalie S Joe^{1

2}, Yuanfeng Wang^{1

3}, Harsh H Oza¹, Inês Godet^{1

4

5}, Nubaira Milki⁴, Gregory J Riggins^{1

6}, Daniele M Gilkes^{1

2

4

5}

Affiliations

¹ Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, Baltimore, MD 21231, USA.
² Cellular and Molecular Medicine Program, The Johns Hopkins University School of Medicine, Baltimore, MD 21231, USA.
³ Biochemistry and Molecular Biology Program, The Johns Hopkins University School of Public Health, Baltimore, MD 21205, USA.
⁴ Department of Chemical and Biomolecular Engineering, The Johns Hopkins University, Baltimore, MD 21218, USA.
⁵ Johns Hopkins Institute for NanoBioTechnology, The Johns Hopkins University, Baltimore, MD 21218, USA.
⁶ Department of Neurosurgery, The Johns Hopkins University School of Medicine, Baltimore, MD 21231, USA.

Abstract

Breast cancer is the most diagnosed cancer in women in the world. Mebendazole (MBZ) has been demonstrated to have preclinical efficacy across multiple cancers, including glioblastoma multiforme, medulloblastoma, colon, breast, pancreatic, and thyroid cancers. MBZ was also well tolerated in a recent phase I clinical trial of adults diagnosed with glioma. The mechanisms of action reported so far for MBZ include tubulin disruption, inhibiting angiogenesis, promoting apoptosis, and maintaining stemness. To elucidate additional mechanisms of action for mebendazole (MBZ), we performed RNA sequencing of three different breast cancer cell lines treated with either MBZ or vehicle control. We compared the top genes downregulated upon MBZ treatment with expression profiles of cells treated with over 15,000 perturbagens using the clue.io online analysis tool. In addition to tubulin inhibitors, the gene expression profile that correlated most with MBZ treatment matched the profile of cells treated with known hypoxia-inducible factor (HIF-1α and -2α) inhibitors. The HIF pathway is the main driver of the cellular response to hypoxia, which occurs in solid tumors. Preclinical data support using HIF inhibitors in combination with standard of care to treat solid tumors. Therefore, we tested the hypothesis that MBZ could inhibit the hypoxia response. Using RNA sequencing and HIF-reporter assays, we demonstrate that MBZ inhibits the transcriptional activity of HIFs in breast cancer cell lines and in mouse models of breast cancer by preventing the induction of HIF-1α, HIF-2α, and HIF-1β protein under hypoxia. Taken together, our results suggest that MBZ treatment has additional therapeutic efficacy in the setting of hypoxia and warrants further consideration as a cancer therapy.

Keywords: HIF1; HIF2; breast cancer; digoxin; hypoxia; mebendazole.

Grants and funding

Work in the Gilkes Lab was supported by The Jayne Koskinas Ted Giovanis Foundation for Health and Policy, The Emerson Collective, and The Allegany Health Network. Work in the Riggins Lab was supported by the Virginia & DK Ludwig Fund for Cancer Research.